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已发表论文
基因纳米聚合体和透明质酸聚电解质多层膜组装介导的可控局部基因递送
Authors Teng W, Wang QM, Chen Y, Huang HZ
Published Date October 2014 Volume 2014:9(1) Pages 5013—5024
DOI http://dx.doi.org/10.2147/IJN.S70952
Received 11 July 2014, Accepted 3 September 2014, Published 29 October 2014
Approved for publication by Professor Thomas J. Webster
Abstract: To explore a spatiotemporally controllable gene delivery system with high efficiency and safety, polyelectrolyte multilayer (PEM) films were constructed on titanium or quartz substrates via layer-by-layer self-assembly technique by using plasmid deoxyribonucleic acid-loaded lipopolysaccharide–amine nanopolymersomes (pNPs) as polycations and hyaluronic acid (HA) as polyanions. pNPs were chosen because they have high transfection efficiency (>95%) in mesenchymal stem cells (MSCs) and induce significant angiogenesis in zebrafish in conventional bolus transfection. The assembly process of PEM films was confirmed by analyses of quartz crystal microbalance with dissipation, X-ray photoelectron spectroscopy, infrared, contact angle, and zeta potential along with atomic force microscopy observation. Quartz crystal microbalance with dissipation analysis reveals that this film grows in an exponential mode, pNPs are the main contributor to the film mass, and the film mass can be modulated in a relatively wide range (1.0–29 µg/cm2) by adjusting the deposition layer number. Atomic force microscopy observation shows that the assembly leads to the formation of a patterned film with three-dimensional tree-like nanostructure, where the branches are composed of beaded chains (pNP beads are strung on HA molecular chains), and the incorporated pNPs keep structure intact. In vitro release experiment shows that plasmid deoxyribonucleic acid can be gradually released from films over 14 days, and the released plasmid deoxyribonucleic acid exists in a complex form. In vitro cell experiments demonstrate that PEM films can enhance the adhesion and proliferation of MSCs and efficiently transfect MSCs in situ in vitro for at least 4 days. Our results suggest that a (pNPs/HA)n system can mediate efficient transfection in stem cells in a spatially and temporally controllable pattern, highlighting its huge potential in local gene therapy.
Keywords: localized gene delivery, layer-by-layer self-assembly, gene-loaded nanopolymersomes, hyaluronic acid, polyelectrolyte multilayer films, mesenchymal stem cells
Keywords: localized gene delivery, layer-by-layer self-assembly, gene-loaded nanopolymersomes, hyaluronic acid, polyelectrolyte multilayer films, mesenchymal stem cells
